APPLICATION OF REAL-TIME PCR TECHNLOGIES FOR ANALYSIS OF JCV AS A HUMAN CANCEROGEN

作者：Tsekov Iliya; Ferdinandov Dilyan; Hristova Svetlana; Stoyanova Daniela; Kacarov Krum; Kalvatchev Zlatko^*

来源：Biotechnology & Biotechnological Equipment, 2011, 25(1): 2262-2265.

DOI：10.5504/BBEQ.2011.0023

• real time PCR
• JC polyomavirus
• brain tumours
• colorectal cancer
• polyp
• oncogenesis
• malignant transformation

Viral agents such as Polyomavirus JC (JCV) raise an increased interest and are discussed as factors for development of several human malignancies. Real time PCR techniques allow better characterization of the studied samples as they possess superior sensitivity when compared to conventional PCR. Also quantitative analysis of the experimental results can be performed and information about the dynamics of the process is determined.
Real time PCR technologies were applied to investigate fresh frozen biopsies from 132 patients for two particular JCV genomic sequences (LT and NCCR) that might be related to malignant transformation. Patients were divided into groups according to their histological type, and included: gliobastoma patients (n=39); astrocytoma patients (n=19); oligodendrogliomas (n=12); colorectal cancer patients (n=44); and patients with polyps (n=18). Colon samples were paired with matched adjacent mucosa. Results showed that viral genomic sequences were present in 22.7% to 61.5% among the different groups. Positive samples had a low starting viral number, as a late threshold cycle that varied from 36 to 49 was observed. The prevalence of the non-coding control region (NCCR) was lower than the prevalence of LT and only 12 of all 70 brain samples were found positive. None of the tested premalignant colonic lesions and cancers was positive, whereas two normal tissues had viral load in the range of 10-1000 starting copies.
The disproportion between the results for LT and NCCR might be suggested to be either due to clonally integrated LT fragments, with loss of genetic material, or changes in the NCCR. This might lead to disturbance in the productive course of the infection and a premise for continuous interaction of viral regulatory proteins with cell molecules that are responsible for the control of the cell cycle may be establish. This subsequently might lead to malignant transformation. As infectious diseases acquire relevance in this process, new tools for cancer management and prevention are gained.

• 出版日期2011-2
• 单位中国人民解放军军事医学科学院